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Scientific Background
ULBP2 (UL16 binding protein) is a glycoprotein related to MHC class I molecules, that belongs to the family of unique length 16 (UL16) binding proteins. It is a stress-induced ligand for the activating NKG2D receptor in NK cells. It contains the MHC class-I-like α1-α2 domains but lacks the α3 region present in MICA/B proteins and uses GPI (glycosylphosphatidylinositol) as anchor to the plasma membrane. Stress-ligands respond to viral-infections, heat-shock or other cellular stress triggers, and are also crucial in immune surveillance. ULBP2 is expressed at low levels in normal tissues but overexpressed in several types of cancer (such as liver, breast, cervical and skin), with its levels linking to the severity of the prognosis. Targeting ULBP2, for example with miR-6071, was shown to reduce tumorigenicity in glioblastoma (GM). The use of ligands, small molecules, or other, to target the levels or action of ULBP2 can prove beneficial for NK-based cancer therapy.
Product Description
HIV-based · VSV-G-pseudotyped · SIN=YesThe ULBP2 Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles ready to transduce nearly all types of mammalian cells, including primary and non-dividing cells. These viruses transduce cells with Macaca fascicularis (also known as crab-eating macaque or cynomolgus monkey) ULBP2 (XP_005552169.1) driven by an EF1A promoter. The lentiviruses also transduce a puromycin selection gene.
Technical Details
| Vector Type | HIV-based, VSV-G-pseudotyped lentiviral vector |
|---|---|
| Payload / Construct | ULBP2 transgene (Macaca fascicularis) |
| Target Antigen / Gene | ULBP2 (Cynomolgus) (NKG2DL2, RAET1H) |
| Signaling / Architecture | N/A (transgene expression) |
| Selection Marker | Puromycin |
| Reporter | None |
| Biosafety Level | BSL-2 |
| SIN Vector | Yes |
| Formulation | The lentivirus particles were produced in HEK293T cells in medium containing 90% DMEM + 10% FBS. |
| Supplied As | Two vials (500 µl x 2) of lentivirus at a titer≥107 TU/ml. The titer will vary with each lot; the exact value is provided with each shipment. |
| Storage | −80°C; avoid repeated freeze-thaw cycles |
| Hazardous Shipping | UN3373 |
Applications
- Expression of ULBP2 in cells of interest.
- Generate stable cell lines expressing cynomolgus ULBP2 (puromycin resistant).
Biosafety & Safety
To generate an ULBP2 stable cell line, remove the growth medium 48 hours after transduction and replace it with fresh growth medium containing the appropriate amount of puromycin (as pre-determined from a killing curve), for antibiotic selection of transduced cells. Visit: bpsbioscience.com/cell-line-faq for guidelines on performing a kill curve. The lentiviruses are produced with a SIN (self-inactivation) lentivector which ensures self-inactivation of the lentiviral construct after transduction and after integration into the genomic DNA of the target cells. None of the HIV genes (gag, pol, rev) will be expressed in the transduced cells, as they are expressed from packaging plasmids lacking the packing signal and are not present in the lentivirus particle. Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a Biosafety Level 2 facility. BPS Bioscience recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions. Troubleshooting Guide: Visit bpsbioscience.com/lentivirus-faq for detailed troubleshooting instructions. For further questions, email support@bpsbioscience.com.
To generate a ULBP2 (Cynomolgus)-expressing stable cell line, transduce your cells of interest at an appropriate MOI. Remove growth medium 48 hours post-transduction and replace with fresh medium containing puromycin at a concentration pre-determined by a kill curve. Continue selection until untransduced control cells are fully eliminated.
BSL-2 facilities are required. The lentiviruses are produced using a SIN (self-inactivating) lentivector design, ensuring self-inactivation after integration. HIV accessory genes are not expressed in transduced cells. Always follow applicable institutional and regulatory biosafety guidelines.
Each shipment contains 500 µl × 2 of lentivirus at ≥10⁷ TU/ml. The exact titer is lot-specific and is reported on the Certificate of Analysis provided with each shipment.
This lentivirus encodes a puromycin resistance gene. Following transduction, stable integrants can be selected by puromycin treatment. Determine the optimal puromycin concentration for your cell line using a kill-curve assay prior to selection.
This VSV-G-pseudotyped lentiviral vector has broad tropism and can transduce most mammalian cell types, including primary cells, non-dividing cells, and established cell lines. Transduction efficiency may vary by cell type and MOI.
Store at −80°C immediately upon receipt. Avoid repeated freeze-thaw cycles to preserve titer. Aliquot into single-use volumes before freezing for best results.
BioHippo offers customization and add-on services for selected products. Options may include:
- Custom formulations: Virus particles can be packaged in alternative formulations upon request (additional fees may apply).
- Custom titers: Higher-titer preparations may be available; contact us for feasibility and pricing.
- Bulk orders: Volume discounts are available for qualifying order quantities.
- Technical support: Our scientific team can assist with protocol optimization and troubleshooting.
For all customization inquiries, please use the contact form or email support@biohippo.com.
